Inbred corn line NPFA4734

ABSTRACT

Basically, this invention provides for an inbred corn line designated NPFA4734, methods for producing a corn plant by crossing plants of the inbred line NPFA4734 with plants of another corn plants. The invention relates to the various parts of inbred NPFA4734 including culturable cells. This invention also relates to methods for introducing transgenic transgenes into inbred corn line NPFA4734 and plants produced by said methods.

This application claims the benefit under Title 35, United States Code,119(e) of U.S. provisional application 61/112,265 filed Nov. 7, 2008 andpursuant to 35 U.S.C. 119(f) giving priority under 35 U.S.C. 119(a)through (c) claims the benefit of US PVP Application No. 201000024,filed Oct. 30, 2009.

FIELD OF THE INVENTION

This invention is in the field of corn breeding, specifically relatingto an inbred corn line designated NPFA4734. This invention also is inthe field of hybrid maize production employing the present inbred.

BACKGROUND OF THE INVENTION

The original maize plant was indigenous to the Western Hemisphere. Theplants were weed like and only through the efforts of early breederswere cultivated crop species developed. The crop cultivated by earlybreeders, like the crop today, could be wind pollinated. The physicaltraits of maize are such that wind pollination results inself-pollination or cross-pollination between plants. Each maize planthas a separate male and female flower that contributes to pollination,the tassel and ear, respectively. Natural pollination occurs when windtransfers pollen from tassel to the silks on the corn ears. This type ofpollination has contributed to the wide variation of maize varietiespresent in the Western Hemisphere.

The development of a planned breeding program for maize only occurred inthe last century. A large part of the development of the maize productinto a profitable agricultural crop was due to the work done by landgrant colleges. Originally, maize was an open pollinated variety havingheterogeneous genotypes. The maize farmer selected uniform ears from theyield of these genotypes and preserved them for planting the nextseason. The result was a field of maize plants that were segregating fora variety of traits. This type of maize selection led to; at most,incremental increases in seed yield.

Large increases in seed yield were due to the work done by land grantcolleges that resulted in the development of numerous hybrid cornvarieties in planned breeding programs. Hybrids were developed frominbreds which were developed by selecting corn lines and selfing theselines for several generations to develop homozygous pure inbred lines.One selected inbred line was emasculated and another selected inbredline pollinated the emasculated inbred to produce hybrid seed F1 on theemasculated inbred line. Emasculation of the inbred usually is done bydetasseling the seed parent; however, emasculation can be done in anumber of ways. For example an inbred could have a male sterility factorwhich would eliminate the need to detassel the inbred.

In the early seventies the hybrid corn industry attempted to introduceCMS (cytoplasmic male sterility) into a number of inbred lines.Unfortunately, the CMS inbreds also introduced some very poor agronomicperformance traits into the hybrid seed which caused farmers concerncausing the maize industry to shy away from CMS material for a couple ofdecades thereafter.

However, in the last 10-15 years a number of different male sterilitysystems for maize have been successfully deployed. The mosttraditionally of these male sterility and/or CMS systems for maizeparallel the CMS type systems that have been routinely used in hybridproduction in sunflower.

In the standard CMS system there are three different maize linesrequired to make the hybrid. First, there is a cytoplasmic male-sterileline usually carrying the CMS or some other form of male sterility. Thisline will be the seed producing parent line. Second, there must be afertile inbred line that is the same or isogenic with the seed producinginbred parent but lacking the trait of male sterility. This is amaintainer line needed to make new inbred seed of the seed producingmale sterile parent. Third there is a different inbred which is fertile,has normal cytoplasm and carries a fertility restoring gene. This lineis called the restorer line in the CMS system. The CMS cytoplasm isinherited from the maternal parent (or the seed producing plant);therefore for the hybrid seed produced on such plant to be fertile thepollen used to fertilize this plant must carry the restorer gene. Thepositive aspect of this is that it allows hybrid seed to be producedwithout the need for detasseling the seed parent. However, this systemdoes require breeding of all three types of lines: 1) male sterile-tocarry the CMS: 2) the maintainer line; and, 3) the line carrying thefertility restorer gene.

In some instances, sterile hybrids are produced and the pollen necessaryfor the formation of grain on these hybrids is supplied by interplantingof fertile inbreds in the field with the sterile hybrids.

Whether the seed producing plant is emasculated due to detasseling orCMS or transgenes, the seed produced by crossing two inbreds in thismanner is hybrid seed. This hybrid seed is F1 hybrid seed. The grainproduced by a plant grown from a F1 hybrid seed is referred to as F2 orgrain. Although, all F1 seed and plants, produced by this hybrid seedproduction system using the same two inbreds should be substantially thesame, all F2 grain produced from the F1 plant will be segregating maizematerial.

The hybrid seed production produces hybrid seed which is heterozygous.The heterozygosis results in hybrid plants, which are robust andvigorous plants. Inbreds on the other hand are mostly homozygous. Thishomozygosity renders the inbred lines less vigorous. Inbred seed can bedifficult to produce since the inbreeding process in corn linesdecreases the vigor. However, when two inbred lines are crossed, thehybrid plant evidences greatly increased vigor and seed yield comparedto open pollinated, segregating maize plants. An important consequenceof the homozygosity and the homogenity of the inbred maize lines is thatall hybrid seed produced from any cross of two such elite lines will bethe same hybrid seed and make the same hybrid plant. Thus the use ofinbreds makes hybrid seed which can be reproduced readily.

The ultimate objective of the commercial maize seed companies is toproduce high yielding, agronomically sound plants that perform well incertain regions or areas of the Corn Belt. To produce these types ofhybrids, the companies must develop inbreds, which carry needed traitsinto the hybrid combination. Hybrids are not often uniformly adapted forthe entire Corn Belt, but most often are specifically adapted forregions of the Corn Belt. Northern regions of the Corn Belt requireshorter season hybrids than do southern regions of the Corn Belt.Hybrids that grow well in Colorado and Nebraska soils may not flourishin richer Illinois and Iowa soils. Thus, a variety of major agronomictraits is important in hybrid combination for the various Corn Beltregions, and has an impact on hybrid performance.

Inbred line development and hybrid testing have been emphasized in thepast half-century in commercial maize production as a means to increasehybrid performance. Inbred development is usually done by pedigreeselection. Pedigree selection can be selection in an F2 populationproduced from a planned cross of two genotypes (often elite inbredlines), or selection of progeny of synthetic varieties, open pollinated,composite, or backcrossed populations. This type of selection iseffective for highly inheritable traits, but other traits, for example,yield requires replicated test crosses at a variety of stages foraccurate selection.

Maize breeders select for a variety of traits in inbreds that impacthybrid performance along with selecting for acceptable parental traits.Such traits include: yield potential in hybrid combination; dry down;maturity; grain moisture at harvest; greensnap; resistance to rootlodging; resistance to stalk lodging; grain quality; disease and insectresistance; ear and plant height. Additionally, Hybrid performance willdiffer in different soil types such as low levels of organic matter,clay, sand, black, high pH, low pH; or in different environments such aswet environments, drought environments, and no tillage conditions. Thesetraits appear to be governed by a complex genetic system that makesselection and breeding of an inbred line extremely difficult. Even if aninbred in hybrid combination has excellent yield (a desiredcharacteristic), it may not be useful because it fails to haveacceptable parental traits such as seed yield, seed size, pollenproduction, good silks, plant height, etc.

To illustrate the difficulty of breeding and developing inbred lines,the following example is given. Two inbreds compared for similarity of29 traits differed significantly for 18 traits between the two lines. If18 simply inherited single gene traits were polymorphic with genefrequencies of 0.5 in the parental lines, and assuming independentsegregation (as would essentially be the case if each trait resided on adifferent chromosome arm), then the specific combination of these traitsas embodied in an inbred would only be expected to become fixed at arate of one in 262,144 possible homozygous genetic combinations.Selection of the specific inbred combination is also influenced by thespecific selection environment on many of these 18 traits which makesthe probability of obtaining this one inbred even more remote. Inaddition, most traits in the corn genome are regrettably not singledominant genes but are multi-genetic with additive gene action notdominant gene action. Thus, the general procedure of producing a nonsegregating F1 generation and self pollinating to produce a F2generation that segregates for traits and selecting progeny with thevisual traits desired does not easily lead to an useful inbred. Greatcare and breeder expertise must be used in selection of breedingmaterial to continue to increase yield and the agronomics of inbreds andresultant commercial hybrids.

Certain regions of the Corn Belt have specific difficulties that otherregions may not have. Thus the hybrids developed from the inbreds haveto have traits that overcome or at least minimize these regional growingproblems. Examples of these problems include in the eastern corn beltGray Leaf Spot, in the north cool temperatures during seedlingemergence, in the Nebraska region CLN (Corn Lethal Necrosis) and in thewest soil that has excessively high pH levels. The industry oftentargets inbreds that address these issues specifically forming nicheproducts. However, the aim of most large seed producers is to provide anumber of traits to each inbred so that the corresponding hybrid can beuseful in broader regions of the Corn Belt. The new biotechnologytechniques such as Microsatellites, RFLPs, RAPDs and the like haveprovided breeders with additional tools to accomplish these goals.

SUMMARY OF THE INVENTION

The present invention relates to an inbred corn line NPFA4734.Specifically, this invention relates to plants and seeds of this line.Additionally, this relates to a method of producing from this inbred,hybrid seed corn and hybrid plants with seeds from such hybrid seed.More particularly, this invention relates to the unique combination oftraits that combine in corn line NPFA4734.

Generally then, broadly the present invention includes an inbred cornseed designated NPFA4734. This seed produces a corn plant.

The invention also includes the tissue culture of regenerable cells ofNPFA4734 wherein the cells of the tissue culture regenerates plantscapable of expressing the genotype of NPFA4734. The tissue culture isselected from the group consisting of leaf, pollen, embryo, root, roottip, guard cell, ovule, seed, anther, silk, flower, kernel, ear, cob,husk and stalk, cell and protoplast thereof. The corn plant regeneratedfrom NPFA4734 or any part thereof is included in the present invention.The present invention includes regenerated corn plants that are capableof expressing NPFA4734's genotype, phenotype or mutants or variantsthereof.

The invention extends to hybrid seed produced by planting, inpollinating proximity which includes using preserved maize pollen asexplained in U.S. Pat. No. 5,596,838 to Greaves, seeds of corn inbredlines NPFA4734 and another inbred line if preserved pollen is not used;cultivating corn plants resulting from said planting; preventing pollenproduction by the plants of one of the inbred lines if two are employed;allowing cross pollination to occur between said inbred lines; andharvesting seeds produced on plants of the selected inbred. The hybridseed produced by hybrid combination of plants of inbred corn seeddesignated NPFA4734 and plants of another inbred line are apart of thepresent invention. This inventions scope covers hybrid plants and theplant parts including the grain and pollen grown from this hybrid seed.

The invention further includes a method of hybrid F1 production. A firstgeneration (F1) hybrid corn plant produced by the process of plantingseeds of corn inbred line NPFA4734; cultivating corn plants resultingfrom said planting; permitting pollen from another inbred line to crosspollinate inbred line NPFA4734; harvesting seeds produced on plants ofthe inbred; and growing a harvested seed are part of the method of thisinvention.

The present invention also encompasses a method of introducing at leastone targeted trait into maize inbred line comprising the steps of: (a)crossing plant grown from the present invention seed which is therecurrent parent, representative seed of which has been deposited, withthe donor plant of another maize line that comprises at least one targettrait selected from the group consisting of male sterility, herbicideresistance, insect resistance, disease resistance, amylose starch, andwaxy starch to produce F1 plants; (b) selecting from the F1 plants thathave at least one of the targeted traits, forming a pool of progenyplants with the targeted trait; (c) crossing the pool of progeny plantswith the present invention which is the recurrent parent to producebackcrossed progeny plants with the targeted trait; (d) selecting forbackcrossed progeny plants that have at least one of the target traitsand physiological and morphological characteristics of maize inbred lineof the recurrent parent, listed in Table 1 forming a pool of selectedbackcrossed progeny plants; and (e) crossing the selected backcrossedprogeny plants to the recurrent parent and selecting from the resultingplants for the targeted trait and physiological and morphologicalcharacteristics of maize inbred line of the recurrent parent, listed inTable 1 and reselecting from the pool of resulting plants and repeatingthe crossing to the recurrent parent and selecting step in succession toform a plant that comprise the desired trait and all of thephysiological and morphological characteristics of maize inbred line ofthe recurrent parent if the present invention listed in Table 1 asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

This method and the following method of introducing traits can be donewith less back crossing events if the trait and/or the genotype of thepresent invention are selected for or identified through the use ofmarkers. SSR, microsatellites, SNP and the like decrease the amount ofbreeding time required to locate a line with the desired trait or traitsand the characteristics of the present invention. Backcrossing in two oreven three traits (for example the glyphosate, Europe corn borer, cornrootworm resistant genes) is routinely done with the use of markerassisted breeding techniques. This introduction of transgenes ormutations into a maize line is often called single gene conversion.Although, presently more than one gene particularly transgenes ormutations which are readily tracked with markers can be moved during thesame “single gene conversion” process, resulting in a line with theaddition of more targeted traits than just the one, but still having thecharacteristics of the present invention plus those characteristicsadded by the targeted traits.

The method of introducing a desired trait into maize inbred linecomprising: (a) crossing plant grown from the present invention seed,representative seed of which has been deposited the recurrent parent,with plant of another maize line that comprises at least one targettrait selected from the group consisting of nucleic acid encoding anenzyme selected from the group consisting of phytase, stearyl-ACPdesaturase, fructosyltransferase, levansucrase, amylase, invertase andstarch branching enzyme, the donor parent to produce F1 plants; (b)selecting for the targeted trait from the F1 plants, forming a pool ofprogeny plants; (c) crossing the progeny plants with the recurrentparent to produce backcrossed progeny plants; (d) selecting forbackcrossed progeny plants that have at least one of the target traitand physiological and morphological characteristics of maize inbred lineof the present invention as listed in Table 1 forming a pool ofbackcrossed progeny plants; and repeating a step of crossing the newpool with the recurrent parent and selecting for the targeted trait andthe recurrent parents characteristics until the selected plant isessentially the recurrent parent with the targeted trait or targetedtraits. This selection and crossing may take at least 4 backcrosses ifmarker assisted breeding is not employed.

The inbred line and seed of the present invention are employed to carrythe agronomic package into the hybrid. Additionally, the inbred line isoften carrying transgenes that are introduced in to the hybrid seed.

Likewise included is a first generation (F1) hybrid corn plant producedby the process of planting seeds of corn inbred line NPFA4734;cultivating corn plants resulting from said planting; permitting pollenfrom inbred line NPFA4734 to cross pollinate another inbred line;harvesting seeds produced on plants of the inbred; and growing a plantfrom such a harvested seed.

A number of different techniques exist which are designed to avoiddetasseling in maize hybrid production. Some examples are switchablemale sterility, lethal genes in the pollen or anther, inducible malesterility, male sterility genes with chemical restorers. There arenumerous patented means of improving upon the hybrid production system.Some examples include U.S. Pat. No. 6,025,546, which relates to the useof tapetum-specific promoters and the barnase gene to produce malesterility; U.S. Pat. No. 6,627,799 relates to modifying stamen cells toprovide male sterility. Therefore, one aspect of the current inventionconcerns the present invention comprising one or more gene(s) capable ofrestoring male fertility to male-sterile maize inbreds or hybrids and/orgenes or traits to produce male sterility in maize inbreds or hybrids.

The inbred corn line NPFA4734 and at least one transgenic gene adaptedto give NPFA4734 additional and/or altered phenotypic traits are withinthe scope of the invention. Such transgenes are usually associated withregulatory elements (promoters, enhancers, terminators and the like).Presently, transgenes provide the invention with traits such as insectresistance, herbicide resistance, disease resistance increased ordeceased starch or sugars or oils, increased or decreased life cycle orother altered trait.

The present invention includes inbred corn line NPFA4734 and at leastone transgenic gene adapted to give NPFA4734 modified starch traits.Furthermore this invention includes the inbred corn line NPFA4734 and atleast one mutant gene adapted to give modified starch, acid or oiltraits, i.e. amylase, waxy, amylose extender or amylose. The presentinvention includes the inbred corn line NPFA4734 and at least onetransgenic gene: bacillus thuringiensis, the bar or pat gene encodingPhosphinothricin acetyl Transferase, Gdha gene, GOX, VIP, EPSP synthasegene, low phytic acid producing gene, and zein. The inbred corn lineNPFA4734 and at least one transgenic gene useful as a selectable markeror a screenable marker is covered by the present invention.

A tissue culture of the regenerable cells of hybrid plants produced withuse of NPFA4734 genetic material is covered by this invention. A tissueculture of the regenerable cells of the corn plant produced by themethod described above is also included.

DEFINITIONS

In the description and examples, which follow, a number of terms areused. In order to provide a clear and consistent understanding of thespecifications and claims, including the scope to be given such terms,the following definitions are provided.

Early Season Trait Codes

Emergence (EMRGR): Recorded when 50% of the plots in the trial are at V1(1 leaf collar) growth stage.

1=All plants have emerged and are uniform in size

3=All plants have emerged but are not completely uniform

5=Most plants have emerged with some just beginning to break the soilsurface, noticeable lack of uniformity

7=Less than 50% of the plants have emerged, and lack of uniformity isvery noticeable

9=A few plants have emerged but most remain under the soil surface.

If growing conditions are good, there may be no differences to note.Take notes only in trials that show hybrid differences.

Seedling Growth (SVGRR): Recorded between V3 and V5 (3-5 leaf stage)giving greatest weight to seedling plant size and secondary weight touniform growth. Take notes only at locations where there are detectabledifferences between hybrids.

1=Large plant size and uniform growth

3=Acceptable plant size and uniform growth

5=Acceptable plant size and might be a little non-uniform

7=Weak looking plants and non-uniform growth

9=Small plants with poor uniformity

When taking emergence notes, hybrid ratings are relative to each otherover the trial. Purpling (PRPLR): Emergence and/or early growth rating.Purpling is more pronounced on the under sides of leaf blades especiallyon midribs.

1=No plants showing purple color

3=30% plants showing purple color

5=50% plants showing purple color

7=70% plants showing purple color

9=90+% plants showing purple color

Herbicide Injury (HRBDR) List the herbicide type, which is being rated.Then rated each hybrid/variety injury as indicated below.

1=No apparent reduction in biomass or other injury symptoms

5=Moderate reduction in biomass with some signs of sensitivity

9=Severe reduction in biomass with some mortality

Mid-Season Traits Codes

Heat Units to 50% Silk (HU5SN): Recorded the day when 50% of all plantswithin a plot show 2 cm or more silk protruding from the ear. Converteddays to accumulated heat units from planting.

Heat units to 50% Pollen Shed (HUPSN): Recorded the day when 50% of allplants within a plot are shedding pollen. Converted days to accumulatedheat units from planting.

Plant Height in cm (ERHTN): After pollination, recorded average plantheight of each plot. Measured from ground to base of leaf node. Three ormore locations recorded.

Plant Ear Height in cm (PLHTN): After pollination, recorded average earheight of each plot. Measured from ground to base of ear node (shank).Three or more locations should be recorded.

Root Lodging Early % (ERTLP): Early root lodging occurs up to about twoweeks after flowering and usually involves goosenecking. Counted thenumber of root lodged plants and converted to percentage. For FieldEvaluation Test plots (FET), recorded lodged plants out of 50 plantsfrom two locations in each hybrid strip, sum, and record percentage.

Foliar Disease (LFDSR): Foliar disease ratings taken one month beforeharvest through harvest. The predominant disease should be listed in thetrial information and individual hybrid ratings should be given. Ratingsare collected if at least one hybrid has a rating of 5 or greater andthere is a spread of 3 in the ratings.

1=No lesions to two lesions per leaf.

3=A few scattered lesions on the leaf. About five to ten percent of theleaf surface is affected.

5=A moderate number of lesions are on the leaf. About 15 to 20 percentof the leaf surface is affected.

7=Abundant lesions are on the leaf. About 30 to 40 percent of the leafsurface is affected.

9=Highly abundant lesions (>50 percent) on the leaf. Lesions are highlycoalesced. Plants may be prematurely killed.

Data collection (as described above) on the following diseases:

Common Rust (CR) Eye Spot (ES) Gray Leaf Spot (GLS) Northern Corn LeafBlight (NCLB) Stewart's Bacterial Wilt (SBW) Southern Corn Leaf Blight(SCLB) Southern Rust (SR) Corn Virus Complex (CVC)Maize response to diseases can also be rated as:R=Resistant=1 to 2 ratingMR=Moderately Resistant=3 to 4 ratingMS=Moderately Susceptible=5 to 6 ratingS=Susceptible=7 to 9 ratingPreharvest Trait Codes

Heat units to Black Layer (HUBLN): Recorded the day when 50% of allplants within a plot reach black layer stage. Converted days toaccumulated heat units from planting. Notes taken on border rows offour-row plots.

Harvest Population (HAVPN): Counted the number of plants in yield rows,excluding tillers, in each plot. For FET plots, count a thousandth of anacre two times and record the average.

Barren Plants (BRRNP): Counted the number of plants in yield rows havingno ears and/or abnormal ears with less than 50 kernels. For FET plots,counted barren plants out of 50 from two locations in each hybrid strip,sum, and record the percentage. Data collected on entire trial.

Dropped Ears (DROPP): Counted the numbers of ears lying on the ground inyield rows. For FET plots, count dropped ears from the area of 50 plantsfrom two locations in each hybrid strip, sum, and record the percentage.

Stalk Lodging % (STKLP): Stalk lodging will be reported as number ofplants broken below the ear without pushing, excluding green snappedplants. Record trials with approximately five percent or more averagestalk lodging. Counted the number of broken plants in yield rows andconverted to percent. For FET plots, counted stalk lodged plants out of50 from two locations in each hybrid strip, sum, and recorded thepercentage.

Root Lodging Late % (LRTLP): Late root lodging can usually start tooccur about two weeks after flowering and involves lodging at the baseof the plant. Plants leaning at a 30-degree angle or more from thevertical are considered lodged. Counted the number of root lodged plantsin yield rows and converted to percent. For FET plots, counted rootlodged plants out of 50 from two locations in each hybrid strip, sum,and record the percentage.

Push Test for Stalk and Root Quality on Erect Plants % (PSTSP): The pushtest is applied to trials with approximately five percent or lessaverage stalk lodging. Plants are pushed that are not root lodged orbroken prior to the push test. Standing next to the plant, the hand isplaced at the top ear and pushed to arm's length. Push one of the borderrows (four-row small plot) into an adjacent plot border row. Counted thenumber of plants leaning at a 30-degree angle or more from the vertical,including plants with broken stalks prior to pushing, did not countplants that have strong rinds that snap rather than bend over easily.For FET plots, push 50 plants from two interior locations of each hybridstrip, sum, and record the percentage. The goal of the push test is toidentify stalk rot and stalk lodging potential, NOT ECB injury. If ECBinjury was present, only did a push test on the ECB trials.

Data may be collected in the following manner:

PUSXN: Push ten plants and enter the number of plants that do not remainupright.

PSTSP: This is a percent. If you push 10 plants you can simply enter 10times the number of plants that do not remain upright (i.e. 2=20) to getthe percentage.

Either method will work.

Intactness (INTLR):

1=Healthy appearance, tops unbroken

5=25% of tops broken

9=majority of tops broken

Plant Appearance (PLTAR): This is a visual rating based on general plantappearance taking into account all factors of intactness, pest, anddisease pressure.

1=Complete plant with healthy appearance

5=plants look OK

9=Plants not acceptable

Green Snap (GRSNP): Counted the number of plants in yield rows thatsnapped below the ear due to brittleness associated with high winds. ForFET plots, count snapped plants out of 50 from two locations in eachhybrid strip, sum, and record the percentage.

Stay-green (STGRP): This is an assessment of the ability of a grainhybrid to retain green color as maturity approaches (taken near the timeof black-layer) and should not be a reflection of hybrid maturity orleaf disease. Recorded % of green tissue. This may be listed as a StayGreen Rating instead of a Percentage.

Stay Green Rating (STGRR): This is an assessment of the ability of agrain hybrid to retain green color as maturity approached (taken nearthe time of black layer or if major differences are noted later). Thisrating should not be a reflection of the hybrid maturity or leafdisease.

1=solid Green Plant

9=no green tissue

Ear/Kernel Rots (KRDSR): If ear or kernel rots are present, husk tenconsecutive ears in each plot and count the number that have evidence ofear or kernel rots, multiply by 10, and round up to the nearest ratingas described below. Identify and recorded the disease primarilyresponsible for the rot.

1=No rots, 0% of the ears infected.

3=Up to 10% of the ears infected.

5=11 to 20% of the ears infected.

7=21 to 35% of the ears infected.

9=36% or more of the ears infected.

Grain Quality (GRQUR): Husked back several ears after black layer stageand observed kernel cap integrity and relative amount of soft starchendosperm along the sides of kernels.

1=smooth kernel caps and or 10% or less soft starch

3=slight kernel wrinkles and or 30% soft starch

7=moderate kernel wrinkles and or 70% soft starch

9=severe kernel wrinkled and or 90% or more soft starch

Preharvest Hybrid Characteristics-Hybrid Owners

Ear Shape: Slender, Semi-Blocky, Blocky

DESHR:

1=Blocky

5=Semi-blocky

9=Slender

Ear Type: Fixed, Semi-Fixed, Flex (Home location: Thin outside row,every other plant for half of row.)

EARFR:

1=Flex

5=Semi-flex

9=Fixed

Husk Cover: Short, Medium, Long

HSKCR:

1=Long

5=Medium

9=Short

Kernel Depth: Shallow, Medium, Deep

KRLNR:

1=Deep

5=Medium

9=Short (shallow)

Shank Length: Short, Medium, Long

SHLNR:

1=Short

5=Medium

9=Long

Cob Color (COBCR):

1=White

5=Pink

9=Dark Red

Kernel Row Number: Enter average of 3 ears (KRRWN): The average numberof kernel rows on 3 ears.

Cob diameter (COBDR): Cob diameter to be taken with template.

1: small

5: Medium

9: Large

Corn: Harvest Trait Codes

Number of Rows Harvested (NRHAN)

Plot Width (RWIDN)

Plot Length (RLENN)

Yield Lb/Plot (YGSMN)

Test Weight in Lb/Bu (TSTWN)

Moisture % (MST_P)

Adjusted Yield in Bu/A (YBUAN)—entered or calculated

Color Codes

A. Kernel Type: (KRTPN)

1) Dent 2) Flint

B. Endosperm Type: (KRTEN)

1) normal 2) amylase 3) waxy 4) other

C. Sterile Type: (MSCT)

1.) no if yes cytoplasm type 2.) c-type 3.) s-type

D. Anthocyanin of Brace Roots: (PBRCC)

The presences of color on 60% of the brace roots during pollen shed.

1) Absent 2) Faint 3) Moderate 4) Dark 5) Brace Roots not present

E. Anther Color: (ANTCC)

(at 50 percent pollen shed newly extruded anthers, pollen not yet shed)

1) Yellow 2) Red 3) Pink 4) Purple

F. Glume Color: (GLMCC)

(color of glumes prior to pollen shed)

1) Red 2) Green

G. Silk Color: (SLKCC)

Taken at a late flowering stage when all plants have fully extrudedsilk.

Silks at least 2″, long but still fresh.

1) Yellow 2) Pink 3) Red

H. Kernel Color: (KERCC)

The main color of the kernel from at least three ears per ear family.

1) Yellow

2) White

I. Cob Color: (COBCC)

The main color of the cob after shelling from at least three ears perear family.

1) Red 2) Pink 3) White

Color Choices:

1. light green 2. medium green 3. dark green 4. very dark green 5.green-yellow 6. pale yellow 7. yellow 8. yelow-orange 9. salmon 10.pink-orange 11. pink 12. light red 13. cherry red 14. red 15. red andwhite 16. pale purple (describe) 17. purple 18. colorless 19. white 20.white capped 21. buff 22. tan 23. brown 24. bronze 25. variegated 26.other (describe)

Form Input # ABR. Description Value A1 EMRGN Final number of plants perplot # A2 REGNN Region Developed: 1.Northwest # 2.Northcentral3.Northeast 4.Southeast 5.Southcentral 6.Southwest 7.Other A3 CRTYNCross type: 1.sc 2.dc 3.3w 4.msc 5.m3w # 6.inbred 7.rel. line 8.other A4KRTPN Kernel type: 1.sweet 2.dent 3.flint 4.flour # 5.pop 6.ornamental7.pipecorn 8.other A5 EMERN Days to Emergence EMERN #Days B1 ERTLP %Root lodging: (before anthesis): #% B2 GRSNP % Brittle snapping: (beforeanthesis): #% C1 TBANN Tassel branch angle of 2nd primary degree lateralbranch (at anthesis): C10 HUPSN Heat units to 50% pollen shed: (from #HUemergence) C11 SLKCN Silk color: #/Munsell value C12 HUPSN Heat units to50% silk: (from emergence) #HU C13 DSAZN Days to 50% silk in adaptedzone: #Days C14 HU9PN Heat units to 90% pollen shed: (from #HUemergence) C15 HU19N Heat units from 10% to 90% pollen shed: #HU C16DA19N Days from 10% to 90% pollen shed: #Days C2 LSPUR Leaf sheathpubescence of second leaf # above the ear (at anthesis) 1-9 (1 = none):C3 ANGBN Angle between stalk and 2nd leaf above degree the ear (atanthesis): C4 CR2LN Color of 2nd leaf above the ear #/Munsell (atanthesis): value C5 GLCRN Glume Color: #/Munsell value C6 GLCBN Glumecolor bars perpendicular to their # veins (glume bands): 1.absent2.present C7 ANTCN Anther color: #/Munsell value C8 PLQUR Pollen Shed:1-9 (0 = male sterile) # C9 HU1PN Heat units to 10% pollen shed: (from#HU emergence) D1 LAERN Number of leaves above the top ear node: # D10LTBRN Number of lateral tassel branches that # originate from thecentral spike: D11 EARPN Number of ears per stalk: # D12 APBRRAnthocyanin pigment of brace roots: # 1.absent 2.faint 3.moderate 4.darkD13 TILLN Number of tillers: # D14 HSKCN Husk color 25 days after 50%silk: (fresh) #/Munsell value D2 MLWVR Leaf marginal waves: 1-9 (1 =none) # D3 LFLCR Leaf longitudinal creases: 1-9 (1 = none) # D4 ERLLNLength of ear leaf at the top ear node: # cm D5 ERLWN Width of ear leafat the top ear node at the # cm widest point: D6 PLHTN Plant height totassel tip: # cm D7 ERHCN Plant height to the top ear node: # cm D8LTEIN Length of the internode between the ear # cm node and the nodeabove: D9 LTASN Length of the tassel from top leaf collar to # cm tasseltip: E1 HSKDN Husk color 65 days after 50% silk: (dry) #/Munsell valueE10 DSGMN Days from 50% silk to 25% grain # Days moisture in adaptedzone: E11 SHLNN Shank length: # cm E12 ERLNN Ear length: # cm E13 ERDINDiameter of the ear at the midpoint: # mm E14 EWGTN Weight of a huskedear: # gm E15 KRRWR Kernel rows: 1.indistinct 2.distinct # E16 KRNARKernel row alignment: 1.straight # 2.slightly curved 3.curved E17 ETAPREar taper: 1.slight 2.average 3.extreme # E18 KRRWN Number of kernelrows: # E19 COBCN Cob color: #/Munsell value E2 HSKTR Husk tightness 65days after 50% silk: # 1-9 (1 = loose) E20 COBDN Diameter of the cob atthe midpoint: # mm E21 YBUAN Yield: # kg/ha E22 KRTEN Endosperm type:1.sweet 2.extra sweet 3 3.normal 4.high amylose 5.waxy 6.high protein7.high lysine 8.super sweet 9.high oil 10.other E23 KRCLN Hard endospermcolor: #/Munsell value E24 ALECN Aleurone color: #/Munsell value E25ALCPR Aleurone color pattern: 1.homozygous # 2.segregating E26 KRLNNKernel length: # mm E27 KRWDN Kernel width: # mm E28 KRDPN Kernelthickness: # mm E29 K1KHN 100 kernel weight: # gm E3 HSKCR Huskextension: 1.short (ear exposed) # 2.medium (8 cm) 3.long (8-10 cm)4.very long (>10 cm) E30 KRPRN % round kernels on 13/64 slotted screen:#% E4 HEPSR Position of ear 65 days after 50% silk: # 1.upright2.horizontal 3.pendent E5 STGRP Staygreen 65 days after anthesis: 1-9 #(1 = worst) E6 DPOPP % dropped ears 65 days after anthesis: % E7 LRTRP %root lodging 65 days after anthesis: % E8 HU25N Heat units to 25% grainmoisture: (from # HU emergence) E9 HUSGN Heat units from 50% silk to 25%grain # HU moisture in adapted zone:

DETAILED DESCRIPTION OF THE INVENTION

The inbred provides uniformity and stability within the limits ofenvironmental influence for traits as described in the VarietyDescription Information (Table 1) that follows.

The inbred has been produced through a dihaploid system or isself-pollinated for a sufficient number of generations to give inbreduniformity. During plant selection in each generation, the uniformity ofplant type was selected to ensure homozygosity and phenotypic stability.The line has been increased in isolated farmland environments with dataon uniformity and agronomic traits being observed to assure uniformityand stability. No variant traits have been observed or are expected inNPFA4734.

The best method of producing the invention is by planting the seed ofNPFA4734 which is substantially homozygous and self-pollinating or sibpollinating the resultant plant in an isolated environment, andharvesting the resultant seed.

TABLE 1 NPFF7405 VARIETY DESCRIPTION INFORMATION  #1 Type: Dent  #2Region Best Adapted: Broadly adapted *MG Group **Maturity Range HybridRM***(estimate) 4 98-102 99  3. MATURITY (In Region Best Adaptability:show Heat Unit Formula in Comments section): DAYS HEAT UNITS 071 1213.7From emergence to 50% of plants in silk 073 1258.7 From emergence to 50%of plants in pollen 003 0062.6 From 10% to 90% pollen shed ND ______From 50% silk to optimum edible quality ND ______ From 50% silk toharvest at 25% moisture  4. PLANT: Standard Deviation Sample Size 247.0cm Plant Height (to tassel tip) 6.8 15 101.8 cm Ear Height (to base oftop ear node) 9.3 15 016.7 cm Length of Top Ear Internode 1.2 3 0.0Average Number of Tillers 0 70 1.0 Average Number of Ears per Stalk 0 153 Anthocyanin of Brace Roots: 1 = Absent 2 = Faint 3 = Moderate 4 = Dark 5. LEAF: 010.2 cm Width of Ear Node Leaf 1.2 15 074.6 cm Length of EarNode Leaf 3.3 15 06 Number of leaves above top ear 0.0 5 070 degreesLeaf Angle — 1 03 (measure from 2nd leaf above ear at anthesis to stalkabove leaf) 3 Leaf Color (Munsell Code 5GY4/4) 4 Leaf Sheath Pubescence3 (Rate on scale from 1 = none to 9 = like peach fuzz) Marginal Waves(Rate on scale from 1 = none to 9 = many) Longitudinal Creases (Rate onscale from 1 = none to 9 = many)  6. TASSEL: 04 Number of PrimaryLateral Branches 1.2 15 085 Branch Angle from Central Spike — 1 37.3 cmTassel Length 3.7 15 (From top leaf collar to tassel tip) 4 Pollen Shed(Rate on Scale from 0 = male sterile to 9 = heavy shed) 05 Anther Color(Munsell Code 2.5GY8/8) 05 Glume Color (Munsell Code 5GY6/6) 2 BarGlumes (Glume Bands): 1 = Absent 2 = Present  7a. EAR (Unhusked Data):17 Silk Color (3 days after emergence) (Munsell code 5RP4/6) 05 FreshHusk Color (25 days after 50% silking) (Munsell code 5GY7/6) 22 Dry HuskColor (65 days after 50% silking) (Munsell code 2.5Y8/4) 1 Position ofEar at Dry Husk Stage: 1 = Upright 2 = Horizontal 3 = Pendent 3 HuskTightness (Rate on scale from 1 = very loose to 9 = very tight) 2 HuskExtension (at harvest): 1 = Short (ears exposed) 2 = Medium (<8 cm) 3 =Long (8-10 cm beyond ear tip) 4 = Very Long (>10 cm)  7b. EAR (HuskedEar Data): 14.3 cm Ear Length 0.96 15 40.0 mm Ear Diameter at mid-point3.0 15 125.1 gm Ear Weight 21.6 15 15 Number of Kernel Rows 1.7 15 2Kernel Rows: 1 = Indistinct 2 = Distinct 2 Row Alignment: 1 = Straight 2= Slightly Curved 3 = Spiral 05.0 cm Shank Length — 1 1 Ear Taper: 1 =Slight 2 = Average 3 = Extreme  8. KERNEL (Dried): 09.6 mm Kernel Length0.66 15 06.6 mm Kernel Width 0.42 15 05.3 mm Kernel Thickness 0.62 1596.0 % Round Kernels (Shape Grade) — 1 ND Aleurone Color Pattern: 1 =Homozygous 2 = Segregating Describe_______________________________________) ND Aleurone Color (Munsell Code—)    Hard Endosperm Color (Munsell Code 2.5Y8/10) 03 Endosperm Type: 1= Sweet (su1) 2 = Extra Sweet (sh2) 3 = Normal Starch 4 = High AmyloseStarch 5 = Waxy Starch 6 = High Protein 7 = High Lysine 8 = Super Sweet(se) 9 = High Oil 10 = Other ______________________________ 20.6 gmWeight per 100 Kernels (unsized sample) — 1  9. COB: 22.5 mm CobDiameter at mid-point 1.8 15 11 Cob Color (Munsell Code 5R7/6)  10.AGRONOMIC TRAITS: 00.0 % Dropped Ears (at 65 days after anthesis) 01.0 %Pre-anthesis Brittle Snapping 00.0 % Pre-anthesis Root Lodging 02.9 %Post-anthesis Root Lodging (at 65 days after anthesis) #11 Plant TraitsAnther Glume Silk BraceRoot Cob Kernel Color Color Color* Color ColorColor NPFA4734 Yellow* green Pink* Moderate Red* Yellow NPH8431 YellowGreen Red/Purple Moderate Red Yellow striped NP2460 Pink Green GreenModerate Pink Yellow • Very light red almost pink *Silk Color taken at alate flowering stage when all plants have fully extruded silk; silks atleast 2″ long but still fresh. Turns purplish. *Anther can also appearlight green, pale yellow *MG = Maturity group **Maturity is the numberof days from planting to physiological maturity (planting to blacklayer) ***RM = relative maturity

The data provided above is often a color. The Munsell code is areference book of color, which is known and used in the industry and bypersons with ordinary skill in the art of plant breeding. The purity andhomozygosity of inbred NPFA4734 is constantly being tracked usingisozyme genotypes.

Isozyme Genotypes for NPFA4734

Isozyme data were generated for inbred corn line NPFA4734 according toprocedures known and published in the art. The data in Table 2 gives theelectrophoresis data on NPFA4734.

TABLE 2 ELECTROPHORESIS RESULTS FOR NPFA4734 Inbred NPFA4734 PGM1 PGM2PGD1 PGD2 IDH1 IDH2 MDH1 MDH2 9 4 3.8 5 4 4 6 3 Inbred NPFA4734 MDH3MDH4 MDH5 MDH6 ACP1 ACP4 PHI1 ADH1 16 12 12 Mm 2 3 4 4

Table 3 show a comparison between NPFA4734 and a comparable inbred.

In Table 3 NPFA4734 has significantly less yield than NP2460. The twoinbreds show significant differences across all heat unit measurementswith the present invention beginning and completing silking and pollenshedding significantly earlier that NP2460.

TABLE 3 PAIRED INBRED COMPARISON DATA HeatUnits HeatUnits Plant EarInbred Yield to P50 to S50 Height Height NPFA4734 114.7 1300.7 1300.475.5 31.8 NP2460 137 1395.1 1423.3 76.8 38.4 Diff 22.3 94.4 115.3 2.88.5 # Expts 6 7 6 4 4 Prob 0.065* 0.001*** 0.001*** 0.421 0.099* *.05 <Prob <= .10 **.01 < Prob <= .05 ***.00 < Prob <= .01

Table 4 shows the GCA (General Combining Ability) estimates of NPFA4734compared with the GCA estimates of the other inbreds. The estimates showthe general combining ability is weighted by the number ofexperiment/location combinations in which the specific hybridcombination occurs. The interpretation of the data for all traits isthat a positive comparison is a practical advantage. A negativecomparison is a practical disadvantage. The general combining ability ofan inbred is clearly evidenced by the results of the general combiningability estimates. This data compares the inbred parent in a number ofhybrid combinations to a group of “checks”. The check data is from ourcompany's and other companies' hybrids which are commercial products andpre-commercial hybrids, which were grown in the same sets and locations.

TABLE 4 %Late Stay %Stalk %Push Root %Early Root %Dropped Final Green%Green Emergence Vigor Heatunits Heatunits Ear Plant Parent1 Parent2 N07N08 N Yield Moisture TestWeight Lodging Test Lodging Lodging Ears Stand% Snap Rating Rating to S50 to P50 Height Height NPFA4734 14 14 −6.3 1.30.74 −2.11 0.38 0 0.54 15 8.75 NPFA4734 16 16 17.3 0.11 −0.01 −0.64−8.89 0 0.38 −5 28.33 NPFA4734 5 5 6.56 0.12 −0.1 0.25 −2.33 −4.67 −14.84.79 NPFA4734 15 15 10.7 0.79 0.18 −1.15 −7.65 0 0.5 0 48.75 NPFA4734 164 20 −5.4 1.45 0.46 −1.74 5.96 0 0.05 1.67 −3.33 NPFA4734 4 4 −12 1.360.71 −3.33 0 −5 3.75 NPFA4734 4 4 1.45 −0.04 −0.3 −2.71 0 NPFA4734 4 4−13 1.19 0.56 −2.71 0 NPFA4734 7 7 3.03 0.3 0.36 −0.17 0.74 10.44 −0.1950 0 18 5.63 18.75 NPFA4734 7 7 −8.1 2.29 1.06 2.03 0.74 −12.29 −1.48 −50 54 −1.88 11.25 NPFA4734 16 16 15.2 −0.99 −0.26 0.88 −10.53 0 0.56 013.33 NPFA4734 16 16 0.23 1.59 0.9 −5.31 1.53 −0.34 −1.25 −6.25 NPFA473416 16 2.73 1 0.59 −3.31 3.03 0.16 13.75 3.75 NPFA4734 3 3 30.9 −0.3 0.790 28.75 −10.3 −18.5 NPFA4734 3 3 19.7 −2.33 −0.22 0.28 −6.88 4.25 17.38NPFA4734 5 5 17.9 0.6 0.17 −10.56 0.74 −0.27 −15 −2.5 13.75 NPFA4734 1616 14.3 −0.92 −0.22 −3.3 2.38 −0.53 8.75 13.75 NPFA4734 23 9 32 −1.6 10.59 6.25 −11.49 4.62 10.1 0.27 0.54 −14.2 0.03 0.08 −8.82 −19.17 −3.093.27 NPFA4734 9 9 −13 1.74 0.68 −1.79 3.98 0.11 0.88 −56.5 7.96 23.58NPFA4734 2 2 14.8 −1.94 −0.48 0 NPFA4734 16 16 −5.8 0.48 0.83 2.06 −5.630 0.02 −8.33 6.67 NPFA4734 7 7 −5.3 0.31 0.31 1.24 11.55 0.14 2.35−23.56 2.96 18.58 NPFA4734 5 5 −7.4 0.25 0.59 1.02 0.2 2.35 −56.5 4.176.67 NPFA4734 16 16 −6.7 1.17 0.83 1.66 4.77 0 0.02 −8.33 −3.33 NPFA47347 7 −9.1 0.34 0.15 −4.41 10.04 0.14 −0.59 18.44 15.46 33.58 NPFA4734 9 9−7 0.15 0.92 0.92 0.29 1.29 −28.66 8.76 11.74 NPFA4734 7 7 1.45 −0.36−0.03 1.24 14.58 0.14 2.35 27.44 5.46 18.58 NPFA4734 7 7 −20 2.57 2.070.57 14.58 0.14 −0.59 −34.56 5.46 11.08 NPFA4734 2 2 −3.1 −1.99 −1.38 0NPFA4734 7 7 −5.2 1.9 0.97 2.19 14.58 0.14 −0.59 −23.56 10.46 3.58NPFA4734 5 5 2.63 0.54 0.38 0 NPFA4734 8 8 −1 1.09 1.39 −2.21 0.13 0.8825.5 20.46 13.58 NPFA4734 7 7 0.9 −0.4 0.19 −9.77 14.58 0.14 9.71 −15.06−2.04 8.58 NPFA4734 15 15 11.9 0.14 0.16 −0.38 0.35 0.17 3.75 −1.25NPFA4734 7 7 −6.5 −1.07 −2.86 2.12 15.9 0.51 −0.17 0.3 14.46 38.23 −4.467.54 NPFA4734 25 25 −4.5 0.88 0.46 −0.6 −4.67 2.54 −1.89 −0.75 −0.43−0.78 −7.83 −14.48 10.56 4.43 NPFA4734 10 10 2.71 0.14 0.44 −4.06 −11.080.34 −0.18 27.34 11.26 26.74 NPFA4734 7 7 −13 0.51 0.55 −0.37 14.58 0.143.82 −15.06 22.96 16.08 NPFA4734 3 3 −6.9 −0.89 −1.07 0.28 33.13 −12.816.38 NPFA4734 6 6 −25 2.55 1.29 −1.53 −0.57 0.17 2.35 −59.56 −7.04−3.92 NPFA4734 6 6 1.59 −1.2 −0.63 2.88 13.07 0.17 0.88 −3.56 15.4621.08 NPFA4734 7 7 −2.6 0.17 0.41 −1.99 −15.72 0.14 9.71 −15.56 15.4621.08 NPFA4734 7 7 6.02 0.55 0.25 −9.7 14.58 0.14 −0.59 −15.06 5.46 1.08NPFA4734 9 9 −5.1 1.52 1.14 0.68 −11.17 0.11 3.82 25.5 27.96 26.08NPFA4734 3 3 23.7 −2.89 0.76 0 28.75 10.75 17.5 NPFA4734 16 16 0.98 0.120.13 1.11 0.95 0 0.06 −8.33 18.33 NPFA4734 9 9 13.7 −0.32 0.25 −0.362.46 0.11 0.88 25.5 25.46 28.58 NPFA4734 7 7 −2.3 3.14 1.59 −9.1 10.040.14 −0.59 −23.56 2.96 1.08 NPFA4734 8 8 −20 1.39 1.48 0.92 0.29 1.29−37.16 −12.7 −2.19 NPFA4734 7 7 −12 1.11 1.21 −5.81 14.58 0.14 −0.59−4.56 7.96 1.08 NPFA4734 7 7 5.27 −0.59 −0.05 −0.37 8.52 0.14 0.88 27.4415.46 28.58 NPFA4734 3 3 2.25 −2.64 −0.19 0 51.75 −7.25 16.5 NPFA4734 77 −2 −0.12 0.59 3.87 −21.78 0.14 2.35 30.94 17.96 13.58 NPFA4734 6 6−6.5 1.62 1.33 2.88 −11.17 0.17 −0.59 −23.56 10.46 1.08 NPFA4734 7 7 −16−1.35 −0.23 −2.79 14.58 0.14 −0.59 18.44 10.46 18.58 NPFA4734 2 2 4.27−3.53 −0.91 0 NPFA4734 7 7 2.66 2.18 1.77 −3.6 14.58 0.14 −0.59 −15.065.46 18.58 NPFA4734 7 7 −10 0.93 0.61 −5.81 14.58 0.14 −0.59 −47.56 5.4613.58 NPFA4734 10 10 −13 2.23 0.2 −3.33 5.58 0.34 1.29 27.34 11.26 14.24NPFA4734 3 3 −2.3 −1.73 −1.36 0 22.63 24.13 20.88 NPFA4734 7 7 1.14 0.230.15 −1.99 −9.66 0.14 −0.59 −15.06 10.46 13.58 NPFA4734 7 7 −19 2.470.95 −13.87 14.58 0.14 −0.59 −4.56 7.96 1.08 NPFA4734 1 1 12.4 −2.78−0.8 0 NPFA4734 8 8 2.41 −0.1 1.18 0.92 0.29 1.29 −28.66 7.34 −2.19NPFA4734 7 7 −12 0.31 0.45 1.24 8.52 0.14 0.88 18.44 2.96 43.58 NPFA473416 7 23 14.6 −0.67 −0.3 −0.74 24.58 −0.45 8.76 −0.17 0.4 20.67 17.56−0.63 10.54 NPFA4734 10 10 −13 3.45 1.71 0.46 10.13 0.34 −0.18 −5.16−1.24 9.24 NPFA4734 7 7 −0.4 1.55 1.11 −7.42 −15.72 0.14 0.88 −4.5617.96 11.08 NPFA4734 2 2 5.72 0.29 0.65 0.89 NPFA4734 7 7 −8 1.68 1.090.5 14.58 0.14 3.82 −15.06 −4.54 23.58 NPFA4734 6 6 −1.1 1.6 0.71 2.8814.58 0.17 −0.59 7.44 0.46 13.58 NPFA4734 7 7 5 1.44 0.93 0.65 8.52 0.142.35 −34.56 27.96 28.58 NPFA4734 3 3 4.2 −0.18 −0.63 0.22 43.5 −5.25 5.5NPFA4734 6 6 −10 1.67 0.99 −0.06 14.58 0.17 −0.59 −34.56 −2.04 1.08NPFA4734 2 2 0.46 −3.4 −0.5 0.73 NPFA4734 9 9 −0.3 1.74 0.48 0.26 −0.570.11 2.35 6.5 7.96 16.08 NPFA4734 7 7 −5.9 −0.2 0.35 2.19 8.52 0.14 0.88−15.06 17.96 26.08 NPFA4734 7 7 8.19 −1.7 −0.21 0.43 11.55 0.14 0.88−4.56 5.46 31.08 NPFA4734 16 16 12.9 0.22 0.19 −4.55 −5.73 0.16 13.758.75 NPFA4734 16 16 3.6 0.66 0.43 1.15 0.73 0.16 8.75 −6.25 NPFA4734 166 22 3.52 1.35 0.41 1.03 7.92 1.03 8.76 0.4 −0.11 20.67 17.56 1.88 7.04NPFA4734 16 16 4.39 0.78 0.9 −6 3.2 −0.09 3.75 −6.25 NPFA4734 16 16 9.91−0.02 0.15 −2.65 1.19 0.16 −6.25 13.75 NPFA4734 16 7 23 7.31 0.08 0.24−0.22 −2.08 2.87 8.76 0.09 0.6 −1.33 17.56 −3.13 10.54 NPFA4734 7 7 9.6−0.09 0.33 −0.3 14.58 0.14 −0.59 −15.06 7.96 3.58 NPFA4734 6 6 −21 1.051.17 −1.53 14.58 0.17 −0.59 −26.06 5.46 11.08 NPFA4734 10 10 −15 0.921.29 0.46 −29.26 0.34 1.29 −18.16 13.76 21.74 NPFA4734 7 7 −14 1.71 0.94−3.02 0.74 −19.87 −1.76 0 0 −5 13.13 16.25 NPFA4734 7 7 −2.9 1.67 0.96−0.07 0.74 −4.71 −0.19 −5 0 18 0.63 6.25 NPFA4734 7 7 −2.5 1.54 0.88−0.81 0.74 18.01 −0.19 −20 0 −5 −6.88 −8.75 NPFA4734 6 6 −6.5 3.02 1.332.67 0.74 −12.29 −0.22 −25 0 −5 −1.88 3.75 NPFA4734 7 7 1.05 2.87 1.261.3 0.74 −27.44 −1.19 −25 0 18 5.63 11.25 NPFA4734 7 7 2.22 0.53 0.37−10.86 2.46 0.14 −0.59 −12.56 7.96 13.58 NPFA4734 7 7 −12 1.64 1.17−12.47 14.58 0.14 −0.59 −23.56 5.46 13.58 NPFA4734 5 5 6.94 1.24 0.33 0NPFA4734 10 10 −9.7 −0.44 0.55 0.46 4.07 0.34 −0.18 −18.16 6.26 26.74NPFA4734 6 6 −13 −0.64 −0.63 −8.01 8.52 0.17 26.38 6.75 −9.5 NPFA4734 22 6.35 −2.66 −1.4 0.89 NPFA4734 16 16 −2.1 0.2 0.53 0.65 2.54 −0.59 3.753.75 NPFA4734 4 4 −6.6 −0.23 −0.35 2.29 0.19 NPFA4734 4 4 −13 0.84 0.34−1.74 0.17 NPFA4734 5 5 −11 1.82 0.58 −2.06 0 0.6 NPFA4734 5 5 −7.4−0.74 −0.21 3.92 0 0.6 NPFA4734 15 15 15.7 0.16 0.18 −7.24 −1.94 0 0.6NPFA4734 16 16 13.3 −0.03 0.18 −2.33 −11.94 0 0.56 5 28.33 NPFA4734 1515 18.4 0.94 0.71 −4.75 −0.41 0 0.6 NPFA4734 16 16 27.8 −1.34 −0.49−3.37 −7.21 0 0.56 −5 28.33 NPFA4734 16 7 23 22.8 −0.25 0.11 −5.15 14.58−8.36 5.73 0 0.59 0.2 43.67 42.56 15 27.83 NPFA4734 16 16 17 −1.34 −0.57−2.27 −2.13 0 0 −5 28.33 NPFA4734 16 16 17.5 0.75 0.44 −4.14 −7.04 00.56 5 28.33 NPFA4734 15 15 17.1 −0.18 0.17 −2.52 −7.96 0 0.6 NPFA473416 7 23 16.6 −1.19 −0.34 1.52 9.17 −1.2 −2.05 0 0.28 −2.5 0.29 17.2515.75 6.19 25.98 NPFA4734 15 15 5.99 0.5 0.64 −8.05 −3.46 0 0.6 NPFA47345 5 6.45 −3.51 −0.84 −3.33 0.18 0 NPFA4734 7 7 2.04 0.48 0.37 −1.01 0.7410.44 −0.19 15 0 18 0.63 16.25 NPFA4734 6 6 7.86 1.38 0.49 −11.5 0.74−0.22 −25 −5 5.63 6.25 NPFA4734 7 7 4.54 1.71 1.2 −4.34 0.74 18.01 −0.19−25 0 18 3.13 13.75 NPFA4734 7 7 2.54 3.7 1.55 −1.1 0.74 2.86 −0.19 −300 −5 −4.38 3.75 NPFA4734 6 6 20.4 0.37 0.27 −4.68 0.74 −0.22 0 −5 25.6331.25 NPFA4734 7 7 16.9 2.47 1 −1.2 0.74 2.86 −0.19 10 0 −5 3.13 26.25NPFA4734 7 7 −1.7 0.57 0.86 −0.72 9.17 4.01 0.34 7.5 0.79 42.25 59.759.38 8.63 NPFA4734 8 8 20.2 −0.99 −0.56 0.34 0.97 0 0.63 NPFA4734 28 287.99 0.78 0.54 0.43 3.89 −5.19 0 0.42 0.08 −0.41 −29.68 −24.36 5.08 5.78NPFA4734 11 11 1.73 1.1 −1.99 0.61 −18.83 −20.82 1.58 0.25 0.07 −4 −27.222.8 24.27 NPFA4734 15 7 22 8.94 0.24 0.61 0.24 −4.17 −4.66 4.01 0 0.18−2.5 0.29 −5.75 15.75 −2.31 5.69 NPFA4734 7 7 −2 1.32 0.91 2.03 −2.21−27.44 −0.19 30 0 18 10.63 18.75 NPFA4734 7 7 −2.5 1.6 0.65 −4.14 0.74−27.44 −0.19 −30 0 −5 −6.88 −1.25 NPFA4734 7 7 12 1.11 0.42 −0.91 0.7418.01 −0.33 −25 0 18 −1.88 11.25 NPFA4734 42 42 −2.4 −0.17 0.12 −1.19−17.04 4.2 0.18 −0.38 0.35 −0.63 56.31 57.72 −6.99 1.87 NPFA4734 3 3−7.4 2.48 1.05 0 −11.25 −9.25 9.5 NPFA4734 1 1 −5.9 2.75 1.02 0 NPFA47342 2 16.2 −0.74 −0.08 −8 −1.5 15.75 9.5 NPFA4734 3 3 −9.4 0.21 −0.19 0NPFA4734 4 4 −2.6 −0.62 −0.17 −3.19 0 NPFA4734 3 3 −23 −1.2 −0.43 0NPFA4734 16 6 22 11.2 −1.16 −0.21 1.05 5.83 −0.43 −0.99 0 −0.81 −2.50.55 −5.75 −9.25 8.69 8.48 NPFA4734 12 12 −8 −1.68 −1 −5 −0.17 5.88NPFA4734 2 2 13.2 −5.9 −0.1 0.73 NPFA4734 4 4 5.27 −1.06 −0.17 −5.830.56 NPFA4734 15 15 −11 0.24 0.14 0 0.76 0.27 0 NPFA4734 4 4 −1.1 2.370.8 −1.53 0 NPFA4734 3 3 7.06 −0.08 −0.61 1.94 1.76 −2.67 NPFA4734 2 2−34 0.73 −0.17 −1.76 −2.35 0 NPFA4734 2 2 −4.7 −2.11 −0.61 −2.48 1.03 0NPFA4734 2 2 −10 −0.12 −0.87 4.12 0.59 0 XR = 1329 2.5 0.43 0.38 −1.59−0.92 −1.13 3.3 −0.17 0.09 −7.73 0.84 0.23 −0.51 −2.65 5.48 5.35 12.49XH = 145 0.58 0.32 0.32 −1.7 2.34 −0.74 3.3 −0.06 0.03 −7.73 0.84 0.26−0.67 0.17 13.43 5 12.62 XT = 9 8.67 0.09 0.18 0.25 5.54 −0.07 5.39 0.050.13 −14.2 −2.5 0.28 0.08 10.07 12.29 2.7 10.67 XR = GCA Estimate:Weighted by Expt XH = GCA Estimate: Weighted by Parent2 XT = Same as XHbut using only those parent2 with two years of data

Table 5 shows the inbred NPFA4734 in hybrid combination, as Hybrid 1, incomparison with another hybrid, which is adapted for the same region ofthe Corn Belt.

When in this hybrid combination, the present inbred NPFA4734 carriessignificantly less yield but also less moisture in comparison to theother commercial hybrid. The test weight for the commercial hybrid andthe present invention is significantly different.

TABLE 5 PAIRED HYBRID COMPARISON DATA PCTER PCTPUS PCTLR Hybrid YieldMoist TWT L PCTSL H L HYBRID 1 172.2 17 57.3 — 6.2 23.3 14.2 Hybrid 2183 18.2 56.4 — 6 13.3 11.2 # Expts 27 27 24 — 15 6 10 Diff 10.8 1.1 0.9— 0.2 10 3 Prob 0.014** 0.001*** 0.002*** 0.899 0.447 0.507 PCTS HybridPCTDE Stand G PCTGS PctBarren Emerge Vigor HYBRID 1 0 63.9 — — — 3.6 3.3Hybrid 2 0 64.4 — — — 3.8 3.3 # Expts 2 29 — — — 8 6 Diff 0 0.5 — — —0.1 0 Prob 0.465 0.685 1 Hybrid HUS50 HUP50 Pltht Earht HYBRID 1 12671258 277.4 119.4 Hybrid 2 1275 1246 275.8 119.6 # Expts 6 6 5 5 Diff 8.312.5 1.6 0.2 Prob 0.175 0.408 0.778 0.974 *.05 < Prob <= .10 **.01 <Prob <= .05 ***.00 < Prob <= .01

This invention also is directed to methods for producing a corn plant bycrossing a first parent corn plant with a second parent corn plantwherein the first or second parent corn plant is an inbred corn plantfrom the line NPFA4734. Further, both first and second parent cornplants can come from the inbred corn line NPFA4734 which produces a selfof the inbred invention. The present invention can be employed in avariety of breeding methods which can be selected depending on the modeof reproduction, the trait, and the condition of the germplasm. Thus,any breeding methods using the inbred corn line NPFA4734 are part ofthis invention: selfing, backcrosses, hybrid production, and crosses topopulations, and haploid by such old and known methods of using KWSinducers lines, Krasnador inducers, stock six material that induceshaploids and anther culturing and the like.

The present invention may be useful as a male-sterile plant. Sterilitycan be produced by pulling or cutting tassels from the plant,detasseling, use of gametocides, use of genetic material to render theplant sterile using a CMS type of genetic control or a nuclear geneticsterility. Male sterility is employed in a hybrid production byeliminating the pollen from the seed producing parent so when inisolation from other pollen source the only available pollen is thatfrom the second male fertile inbred planted most often in rows near themale sterile inbred.

Methods for genetic male sterility are disclosed in EPO 89/3010153.8, WO90/08828, U.S. Pat. Nos. 4,654,465, 4,727,219, 3,861,709, 5,432,068 and3,710,511. Gametocides which are chemicals or substances that negativelyaffect the pollen or at least the fertility of the pollen can beemployed to provide male sterility.

Unfortunately, for hybrid production nature complicates male sterilityand as a result there are self pollinated female inbred seeds in somehybrid production. Great measures are taken to avoid this inbredproduction in a hybrid field but it unfortunately does occur. If ahybrid bag of seed is tested with molecular markers it may be possibleto detect inbred seed. If the hybrid seed is planted these inbred plantstend to be readily identifiable as runt like appearance—shorter plant,small ear, or other characteristics when the hybrid seed in a bag isplanted. Self pollination of plants grown from these female inbreds seedproduces the female inbred seed. This seed is not sold to the growersfor breeding but only for use as grain and forage.

Process for producing seed comprises planting a group of seed comprisingseed from a hybrid production, one of whose parents is the presentinvention said group, growing plants from this seed, identifying anyinbred plants, selecting and pollinating the inbred plant.

A number of well known methods can be employed to identify the genotypeof maize. The ability to understand the genotype of the presentinvention increases as the technology moves toward better markers foridentifying different components within the maize genetic material. Oneof the oldest methods is the use of isozymes which provides ageneralized footprint of the material. Other markers that were adaptedto provide a higher definition profile include Restriction FragmentLength Polymorphisms (RFLPs), Amplified Fragment Length Polymorphisms(AFLPs), Random Amplified Polymorphic DNAs (RAPDs), Polymerase ChainReaction (there are different types of primers or probes) (PCR),Microsattelites (SSRs), and Single Nucleotide Polymorphisms (SNPs) justto list a few. The use of these and a number of other markers forgathering genotype information is well understood in the industry andcan be found in college textbooks such as Breeding Field Crops, Miltonet. al. Iowa State University Press.

The profile of the inbred of this invention should be close tohomozygous for alleles. A marker profile produced with any of the locusidentifying systems known in the industry will identify a particularallele at particular loci. A F1 hybrid made from the inbred of thisinvention will comprise a marker profile of the sum of both of itsinbred parents. At each locus the allele for the present invention andthe allele for the other inbred parent should be present. Thus theprofile of the present invention will permit identification of hybridsas containing the inbred parent of the present invention. To identifythe female portion of the hybrid the material from the pericarp which ismaternally inherited is employed. The comparison of this maternalprofile with the hybrid profile will allow identification of thepaternal profile. The present invention includes a maize cell that ispart of an inbred or hybrid plant which includes its seed or plant partthat has the marker profile of alleles of the present invention.

Marker systems are not just useful for identification of the presentinvention; they are also useful for breeding and trait conversiontechniques. Polymorphisms in maize permit the use of markers for linkageanalysis. If SSR are employed with flanking primers PCR can be used andSouthern Blots can often be eliminated. Use of flanking markers and PCRand amplification of the material is well known by the industry. Primersfor SSRS and mapping information are publicly available through the helpof the USDA at Maize GDB on the web.

Marker profiles of this invention can identify essentially derivedvarieties or progeny developed with the inbred in its ancestry. Thisinbred may have progeny identified by having a molecular marker profileof at least 25%, to 40%, 45%, 50% to 80% (which includes each of thenumber between these two percentages), 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% geneticcontribution of the present inbred invention, as measured by eitherpercent identity or percent similarity.

The present invention may have a new locus or trait introgressed throughdirect transformation or backcrossing or marker assisted breeding. Abackcross conversion or locus conversion both refer to a product of abackcrossing program.

DNA sequences are introduced through backcrossing (Hallauer et al. inCorn and Corn Improvement, Sprague and Dudley, Third Ed. 1998), withPH8JV utilized as the recurrent parent.

When the present inbred is used as a recurrent parent in a breedingprogram it is often referred to as backcrossing. Backcrossing is oftenemployed to introgression a desired trait or trait(s), either transgenicor nontransgenic, into the recurrent parent. A plant may be selectedwith the trait or the desired locus in one or more backcrosses. Ifmarkers are employed to assist in selection the number of backcrossesneeded to recover the recurrent parent with the desired trait or locuscan be relatively few two or three. However, 3, 4, 5 or more backcrossesare often required to produce the desired inbred with the gene or lociconversion in place. The number of backcrosses needed for a traitintrogression is often linked to the genetics of the trait. Multigenictraits, recessive alleles, unlinked traits, how the traits are inheritedall will play a role in the number of backcrosses that may be necessaryto achieve the desired backcross conversion of the inbred.

Dominant, single gene traits or traits with obvious phenotypic changesare particularly well managed in a backcrossing program. Prior totransformation and prior to markers backcrossing was employed since atleast the 1950's to alter grain color, to move mutations intoinbreds—such as sugary 2, waxy, amylose extender, dull, brittle,shrunken, sugary 1, waxy (wx), shrunken-2,

In a book written by Hallauer entitled Corn and Corn Improvement,Sprague and Dudley, 3rd Ed. 1998 the basics of this type of crossingalong with a number of other corn breeding methods such as recurrent orbulk or mass selection, pedigree breeding, open pollination breeding,marker assisted selection, double haploids development and breeding ataught. The ordinary corn breeder understands these breeding systems andhow to apply them to the present invention therefore repetition of thesebreeding methods need not be listed within this application.

The backcrossing program is more complicated when the trait is arecessive gene. To determine the presence of the recessive gene oftenrequires the use of additionally testing to determine if the trait I hasbeen transferred. Use of markers to detect the gene reduces thecomplexity of trait identification in the progeny. A marker that is aSNP specific for the trait itself can be very useful in increasing theefficiency and speed of tracking a recessive trait within a backcrossingprogram. Backcrossing of recessive traits such as has been preformedsince at least the 1950 as mutant corn was frequently moved into moreelite germplasm. Mutations can be induced in germplasm by the plantbreeder. Mutations can result from plant or seed or pollen exposure totemperature alterations, culturing, radiation in various forms, chemicalmutagens like EMS and others. Some of the mutant genes which have beenidentified in maize include the genotypes: waxy (wx), amylose extender(ae), dull (du), horny (h), shrunken (sh), brittle (bt), floury (fl),opaque (O), and sugary (su). Nomenclature for mutant genes is based onthe effect these mutant genes have on the physical appearance,phenotype, of the kernel. It is also known that within these genotypesthere are genes which produce starch with markedly different functionalproperties even though the phenotypes are the same. Such subspecies havegenerally been given a number after the named genotype, for example,sugary-1), sugary-2 (su2); shrunken 1 and shrunken 2. Traits such as Ht,waxy, shrunken, amylose extender, opaque, sugary 1, 2, dull, IT, IR,sterility, fertility, phytic acid, NLB, SLB, and the like have all beenintrogressed into elite inbreds through backcrossing programs. The lastbackcross generation is usually selfed if necessary to recover theinbred of interest with the introgressed trait.

All plants and plant cells produced using inbred corn line NPFA4734 arewithin the scope of this invention. The invention encompasses the inbredcorn line used in crosses with other, different, corn inbreds to produce(F1) corn hybrid seeds and hybrid plants and the grain produced on thehybrid plant. This invention includes plant and plant cells, which upongrowth and differentiation produce corn plants having the physiologicaland morphological characteristics of the inbred line NPFA4734.

Additionally, this maize can, within the scope of the invention,contain: a mutant gene such as, but not limited to, amylose, amylase,sugary 1, shrunken 1, waxy, AE (amylose extender), dull, brown midrib,or imazethapyr tolerant (IT or IR TM).

This invention also includes transforming of introgressed transgenicgenes, or specific locus into the present invention. The prior art hasan extended list of transgenes, and of specific locus that carrydesirable traits. The transgenes that can be introgressed include butare not limited to insect resistant genes such as Corn Rootworm gene(s)in the event DAS-59122-7, Mir603 Modified Cry3A event, MON 89034, MON88017 Bacillus thuringiensis (Cry genes) Cry34/35Ab1, Cry1A.105, POCry1F, Cry2Ab2, Cry1A, Cry1AB, Cry1Ac Cry3Bb1, or herbicide resistantgenes such as Pat gene or Bar gene, EPSP, the altered protoporphyrinogenoxidase (protox enzyme) U.S. Pat. Nos. 5,767,373, 6,282,837, WO01/12825, or disease resistant genes such as the Mosaic virus resistantgene, etc., or trait altering genes such as lignin genes, floweringgenes, oil modifying genes, senescence genes and the like.

The present invention also encompasses the addition of traits that focuson products or by products of the corn plant such as the sugars, oils,protein, ethanol, biomass and the like. The present invention caninclude a trait that forms an altered carbohydrate or altered starch. Analtered carbohydrate or altered starch be formed by an introgressedgene(s) that affect the synthase, branching enzymes, pullanases,debranching enzymes, isoamylases, alpha amylases, beta amylases, AGP,ADP and other enzymes which effect the amylose, and or amylopectin ratioor content or the branching pattern of starch. The fatty acid modifyinggenes if introgressed into the present invention can also affect starchcontent. Additionally, introgressed genes that are associated with oreffect the starch and carbohydrates can be adapted so that the gene orits enzyme does not necessarily alter the form or formation of thestarch or carbohydrate of the seed or plant; instead the introgressedgene or its RNA, polypeptide, protein or enzyme adapted to degrade,alter, or otherwise change the formed starch or carbohydrate. An exampleof use of an alpha amylase adapted in this manner in maize is shown inU.S. Pat. No. 7,407,677 which is incorporated by reference.

The methods and techniques for inserting, or producing and/oridentifying a mutation or making or reshuffling a transgene andintrogressing the trait or gene into the present invention throughbreeding, transformation, mutating and the like are well known andunderstood by those of ordinary skill in the art.

A number of different inventions exist which are designed to avoiddetasseling in maize hybrid production. Some examples are switchablemale sterility, lethal genes in the pollen or anther, inducible malesterility, male sterility genes with chemical restorers, sterility geneslinked with a parent. U.S. Pat. No. 6,025,546, relates to the use oftapetum-specific promoters and the barnase gene. U.S. Pat. No. 6,627,799relates to modifying stamen cells to provide male sterility. Therefore,one aspect of the current invention concerns the present inventioncomprising one or more gene(s) capable of restoring male fertility tomale-sterile maize inbreds or hybrids.

Various techniques for breeding, moving or altering genetic materialwithin or into the present invention (whether it is an inbred or inhybrid combination) are also known to those skilled in the art. Thesetechniques to list only a few are anther culturing, haploid/doublehaploid production, (stock six, which is a breeding/selection methodusing color markers and is a method that has been in use for forty yearsand is well known to those with skill in the art), transformation,irradiation to produce mutations, chemical or biological mutation agentsand a host of other methods are within the scope of the invention. Allparts of the NPFA4734 plant including its plant cells produced using theinbred corn line is within the scope of this invention. The termtransgenic plant refers to plants having genetic sequences, which areintroduced into the genome of a plant by a transformation method and theprogeny thereof. Transformation methods are means for integrating newgenetic coding sequences into the plant's genome by the incorporation ofthese sequences into a plant through man's assistance, but not bybreeding practices. The transgene once introduced into plant materialand integrated stably can be moved into other germplasm by standardbreeding practices.

Though there are a large number of known methods to transform plants,certain types of plants are more amenable to transformation than areothers. Transformation of dicots is usually achievable for example,tobacco is a readily transformable plant. Monocots can present sometransformation challenges, however, the basic steps of transformingplants monocots have been known in the art for about 15 years. The mostcommon method of maize transformation is referred to as gunning ormicroprojectile bombardment though other methods can be used. Theprocess employs small gold-coated particles coated with DNA which areshot into the transformable material. Detailed techniques for gunningDNA into cells, tissue, callus, embryos, and the like are well known inthe prior art. One example of steps that can be involved in monocottransformation are concisely outlined in U.S. Pat. No. 5,484,956“Fertile Transgenic Zea mays Plants Comprising Heterologous DNA EncodingBacillus Thuringiensis Endotoxin” issued Jan. 16, 1996 and also in U.S.Pat. No. 5,489,520 “Process of Producing Fertile Zea mays Plants andProgeny Comprising a Gene Encoding Phosphinothricin Acetyl Transferase”issued Feb. 6, 1996.

Plant cells such as maize can be transformed not only by the use of agunning device but also by a number of different techniques. Therecombinant DNA molecules of the invention can be introduced into theplant cell in a number of art-recognized ways. Those skilled in the artwill appreciate that the choice of method might depend on the type ofplant, i.e. monocot or dicot, targeted for transformation. Suitablemethods of transforming plant cells include microinjection (Crossway etal., BioTechniques 4:320-334 (1986)), electroporation (Riggs et al,Proc. Natl. Acad. Sci. USA 83:5602-5606 (1986), Agrobacterium mediatedtransformation (Hinchee et al., Biotechnology 6:915-921 (1988)), directgene transfer (Paszkowski et al., EMBO J. 3:2717-2722 (1984)), ballisticparticle acceleration using devices available from Agracetus, Inc.,Madison, Wis. and Dupont, Inc., Wilmington, Del. (see, for example,Sanford et al., U.S. Pat. No. 4,945,050; and McCabe et al.,Biotechnology 6:923-926 (1988)), protoplast transformation/regenerationmethods (see U.S. Pat. No. 5,350,689 issued Sep. 27, 1994 to Ciba-GeigyCorp.), Whiskers technology (See U.S. Pat. Nos. 5,464,765 and 5,302,523)and pollen transformation (see U.S. Pat. No. 5,629,183). Also see,Weissinger et al., Annual Rev. Genet. 22:421-477 (1988); Sanford et al.,Particulate Science and Technology 5:27-37 (1987)(onion); Christou etal., Plant Physiol. 87:671-674 (1988)(soybean); McCabe et al.,Bio/Technology 6:923-926 (1988)(soybean); Datta et al., Bio/Technology8:736-740 (1990) (rice); Klein et al., Proc. Natl. Acad. Sci. USA,85:4305-4309 (1988)(maize); Klein et al., Bio/Technology 6:559-563(1988)(maize); Klein et al., Plant Physiol. 91:440-444 (1988)(maize);Fromm et al., Bio/Technology 8:833-839 (1990); Gordon-Kamm et al., PlantCell 2:603-618 (1990) (maize); and U.S. Pat. Nos. 5,591,616 and5,679,558 (rice).

A further subject of the present invention are the plants which comprisetransformed cells, in particular the plants regenerated from transformedcells. Regeneration is effected by any suitable process, which dependson the nature of the species as described, for example, in thereferences hereinabove. Patents and patent applications which are citedin particular for the processes for transforming plant cells andregenerating plants are the following: U.S. Pat. Nos. 4,459,355,4,536,475, 5,464,763, 5,177,010, 5,187,073, EP 267,159, EP 604 662, EP672 752, U.S. Pat. Nos. 4,945,050, 5,036,006, 5,100,792, 5,371,014,5,478,744, 5,179,022, 5,565,346, 5,484,956, 5,508,468, 5,538,877,5,554,798, 5,489,520, 5,510,318, 5,204,253, 5,405,765, EP 442 174, EP486 233, EP 486 234, EP 539 563, EP 674 725, WO 91/02071 and WO95/06128.

The use of pollen, cotyledons, zygotic embryos, meristems and ovum asthe target issue can eliminate the need for extensive tissue culturework. Generally, cells derived from meristematic tissue are useful. Themethod of transformation of meristematic cells of cereal is taught inthe PCT application WO96/04392. Any number of various cell lines,tissues, calli and plant parts can and have been transformed by thosehaving knowledge in the art. Methods of preparing callus or protoplastsfrom various plants are well known in the art and specific methods aredetailed in patents and references used by those skilled in the art.Cultures can be initiated from most of the above-identified tissue. Theonly true requirement of the transforming plant material is that it canultimately be used to form a transformed plant.

Heterologous means of different natural origin or represents anon-natural state. A host cell transformed with a nucleotide sequencederived from another organism, particularly from another species, thatnucleotide sequence is heterologous with respect to that host cell anddescendants. Heterologous refers to a nucleotide sequence derived fromand inserted into the same natural, original cell type, but which ispresent in a non-natural state, e.g. a different copy number, or underthe control of different regulatory sequences. A transforming nucleotidesequence may comprise a heterologous coding sequence, or heterologousregulatory sequences. Alternatively, the transforming nucleotidesequence may be completely heterologous or may comprise any possiblecombination of heterologous and endogenous nucleic acid sequences.

The DNA used for transformation of these plants clearly may be circular,linear, and double or single stranded. Usually, the DNA is in the formof a plasmid. The plasmid usually contains regulatory and/or targetingsequences which assists the expression or targeting of the gene in theplant. The methods of forming plasmids for transformation are known inthe art. Plasmid components can include such items as: leader sequences,transit polypeptides, promoters, terminators, genes, introns, markergenes, etc. The structures of the gene orientations can be sense,antisense, partial antisense, or partial sense: multiple gene copies canbe used. The transgenic gene can come from various non-plant genes (suchas; bacteria, yeast, animals, and viruses) along with being from plants.

The regulatory promoters employed can be constitutive such as CaMv35S(usually for dicots) and polyubiquitin for monocots or tissue specificpromoters such as CAB promoters, MR7 described in U.S. Pat. No.5,837,848, etc. The prior art promoters, includes but is not limited to,octopine synthase, nopaline synthase, CaMv19S, mannopine synthase. Theseregulatory sequences can be combined with introns, terminators,enhancers, leader sequences and the like in the material used fortransformation.

The isolated DNA is then transformed into the plant. A transgeneintrogressed into this invention typically comprises a nucleotidesequence whose expression is responsible or contributes to the traitunder the control of a promoter appropriate for the expression of thenucleotide sequence at the desired time in the desired tissue or part ofthe plant. Constitutive or inducible promoters are used. The transgenemay also comprise other regulatory elements such as for exampletranslation enhancers or termination signals. In an embodiment, thenucleotide sequence is the coding sequence of a gene and is transcribedand translated into a protein. In another embodiment, the nucleotidesequence encodes an antisense RNA, a sense RNA that is not translated oronly partially translated, a t-RNA, a r-RNA or a sn-RNA.

Where more than one trait are introgressed into inbred invention, it isthat the specific genes are all located at the same genomic locus in thedonor, non-recurrent parent, preferably, in the case of transgenes, aspart of a single DNA construct integrated into the donor's genome.Alternatively, if the genes are located at different genomic loci in thedonor, non-recurrent parent, backcrossing allows torecover all of themorphological and physiological characteristics of the invention inaddition to the multiple genes in the resulting maize inbred line.

The genes responsible for a specific gene trait are generally inheritedthrough the nucleus. Known exceptions are, e.g. the genes for malesterility, some of which are inherited cytoplasmically, but still act assingle gene traits. In an embodiment, a heterologous transgene to betransferred to present invention is integrated into the nuclear genomeof the donor, non-recurrent parent. In another embodiment, aheterologous transgene to be transferred to into present invention isintegrated into the plastid genome of the donor, non-recurrent parent.In an embodiment, a plastid transgene comprises one gene transcribedfrom a single promoter or two or more genes transcribed from a singlepromoter.

In an embodiment, a transgene whose expression results or contributes toa desired trait to be transferred to the present invention comprises avirus resistance trait such as, for example, a MDMV strain B coatprotein gene whose expression confers resistance to mixed infections ofmaize dwarf mosaic virus and maize chlorotic mottle virus in transgenicmaize plants (Murry et al. Biotechnology (1993) 11:1559 64). In anotherembodiment, a transgene comprises a gene encoding an insecticidalprotein, such as, for example, a crystal protein of Bacillusthuringiensis or a vegetative insecticidal protein from Bacillus cereus,such as VIP3 (see for example Estruch et al. Nat Biotechnol (1997)15:137 41). Also see, U.S. Pat. Nos. 5,877,012, 6,291,156; 6,107,2796,291,156 and 6,429,360. In another embodiment, an insecticidal geneintroduced into present invention is a Cry1Ab gene or a portion thereof,for example introgressed into present invention from a maize linecomprising a Bt-11 event as described in U.S. Pat. No. 6,114,608, whichis incorporated herein by reference, or from a maize line comprising a176 event as described in Koziel et al. (1993) Biotechnology 11: 194200. In yet another embodiment, a transgene introgressed into presentinvention comprises a herbicide tolerance gene. For example, expressionof an altered acetohydroxyacid synthase (AHAS) enzyme confers uponplants tolerance to various imidazolinone or sulfonamide herbicides(U.S. Pat. No. 4,761,373). In another embodiment, a non-transgenic traitconferring tolerance to imidazolinones is introgressed into presentinvention (e.g. a “IT” or “IR” trait). U.S. Pat. No. 4,975,374,incorporated herein by reference, relates to plant cells and plantscontaining a gene encoding a mutant glutamine synthetase (GS) resistantto inhibition by herbicides that are known to inhibit GS, e.g.phosphinothricin and methionine sulfoximine. Also, expression of aStreptomyces bar gene encoding a phosphinothricin acetyl transferase inmaize plants results in tolerance to the herbicide phosphinothricin orglufosinate (U.S. Pat. No. 5,489,520). U.S. Pat. No. 5,013,659, which isincorporated herein by reference, is directed to plants that express amutant acetolactate synthase (ALS) that renders the plants resistant toinhibition by sulfonylurea herbicides. U.S. Pat. No. 5,162,602 disclosesplants tolerant to inhibition by cyclohexanedione andaryloxyphenoxypropanoic acid herbicides. The tolerance is conferred byan altered acetyl coenzyme A carboxylase (ACCase). U.S. Pat. No.5,554,798 discloses transgenic glyphosate tolerant maize plants, whichtolerance is conferred by an altered 5-enolpyruvyl-3-phosphoshikimate(EPSP) synthase gene. U.S. Pat. No. 5,804,425 discloses transgenicglyphosate tolerant maize plants, which tolerance is conferred by anEPSP synthase gene derived from Agrobacterium tumefaciens CP-4 strain.Also, tolerance to a protoporphyrinogen oxidase inhibitor is achieved byexpression of a tolerant protoporphyrinogen oxidase enzyme in plants(U.S. Pat. No. 5,767,373). Another trait transferable to the presentinvention confers a safening effect or additional tolerance to aninhibitor of the enzyme hydroxyphenylpyruvate dioxygenase (HPPD) andtransgenes conferring such trait are, for example, described in WO9638567, WO 9802562, WO 9923886, WO 9925842, WO 9749816, WO 9804685 andWO 9904021. All issued patents referred to herein are, in theirentirety, expressly incorporated herein by reference.

In an embodiment, a transgene transferred to present invention comprisesa gene conferring tolerance to a herbicide and at least anothernucleotide sequence encoding another trait, such as for example, aninsecticidal protein. Such combination of single gene traits is forexample a Cry1Ab gene and a bar gene.

By way of example only, specific events (followed by their APHISpetition numbers) that can be transformed or introgressed into maizeplants include the glyphosate tolerant event GA21 (97-09901p) or theglyphosate tolerant event NK603 (00-011-01p), the glyphosatetolerant/Lepidopteran insect resistant event MON 802 (96-31701p) Mon810,Lepidopteran insect resistant event DBT418 (96-29101p), male sterileevent MS3 (95-22801p), Lepidopteran insect resistant event Bt11(95-19501p), phosphinothricin tolerant event B16 (95-14501p),Lepidopteran insect resistant event MON 80100 (95-09301p) and MON 863(01-137-01p), phosphinothricin tolerant events T14, T25 (94-35701p),Lepidopteran insect resistant event 176 (94-31901p) and Western cornrootworm (04-362-01p), and the phosphinothricin tolerant andLepidopteran insect resistant event CBH-351 (92-265-01p). transgeniccorn event designated 3272 taught in US application publication20060230473 (hereby incorporated by reference).

After the transformation of the plant material is complete, the nextstep is identifying the cells or material, which has been transformed.In some cases, a screenable marker is employed such as thebeta-glucuronidase gene of the uidA locus of E. coli. Then, thetransformed cells expressing the colored protein are selected. In manycases, a selectable marker identifies the transformed material. Theputatively transformed material is exposed to a toxic agent at varyingconcentrations. The cells not transformed with the selectable marker,which provides resistance to this toxic agent, die.

Cells or tissues containing the resistant selectable marker generallyproliferate. It has been noted that although selectable markers protectthe cells from some of the toxic affects of the herbicide or antibiotic,the cells may still be slightly affected by the toxic agent by havingslower growth rates. If the transformed material was cell lines thenthese lines are regenerated into plants. The cells' lines are treated toinduce tissue differentiation. Methods of regeneration of cellular maizematerial are well known in the art.

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which maize plants can beregenerated, plant calli, plant clumps, and plant cells that are intactin plants or parts of plants, such as embryos, pollen, ovules, flowers,kernels, ears, cobs, leaves, husks, stalks, roots, root tips, anthers,silk, seeds and the like.

Duncan, Williams, Zehr, and Widholm, Planta (1985) 165:322 332 reflectsthat 97% of the plants cultured that produced callus were capable ofplant regeneration. Subsequent experiments with both inbreds and hybridsproduced 91% regenerable callus that produced plants. In a further studyin 1988, Songstad, Duncan & Widholm in Plant Cell Reports (1988), 7:262265 reports several media additions that enhance regenerability ofcallus of two inbred lines. Other published reports also indicated that“nontraditional” tissues are capable of producing somatic embryogenesisand plant regeneration. K. P. Rao, et al., Maize Genetics CooperationNewsletter, 60:64 65 (1986), refers to somatic embryogenesis from glumecallus cultures and B. V. Conger, et al., Plant Cell Reports, 6:345 347(1987) indicates somatic embryogenesis from the tissue cultures of maizeleaf segments. Thus, it is clear from the literature that the state ofthe art is such that these methods of obtaining plants are, and were,“conventional” in the sense that they are routinely used and have a veryhigh rate of success.

Tissue culture procedures of maize are described in Green and Rhodes,“Plant Regeneration in Tissue Culture of Maize,” Maize for BiologicalResearch (Plant Molecular Biology Association, Charlottesville, Va.1982, at 367 372) and in Duncan, et al., “The Production of CallusCapable of Plant Regeneration from Immature Embryos of Numerous Zea maysGenotypes,” 165 Planta 322 332 (1985). Thus, another aspect of thisinvention is to provide cells that upon growth and differentiationproduce maize plants having the physiological and morphologicalcharacteristics of the present invention. In a preferred embodiment,cells of the present invention are transformed genetically, for examplewith one or more genes described above, for example by using atransformation method described in U.S. Pat. No. 6,114,608, andtransgenic plants of the present invention are obtained and used for theproduction of hybrid maize plants.

The introgression of a Bt11 event into a maize line, such as presentinvention, by backcrossing is exemplified in U.S. Pat. No. 6,114,608,and the present invention is directed to methods of introgressing a Bt11event into present invention and to progeny thereof using for examplethe markers described in U.S. Pat. No. 6,114,608.

Direct selection may be applied where the trait acts as a dominanttrait. An example of a dominant trait is herbicide tolerance. For thisselection process, the progeny of the initial cross are sprayed with theherbicide prior to the backcrossing. The spraying eliminates any plantthat does not have the desired herbicide tolerance characteristic, andonly those plants that have the herbicide tolerance gene are used in thesubsequent backcross. This process is then repeated for the additionalbackcross generations.

Maize is used as human food, livestock feed, and as raw material inindustry. Sweet corn kernels having a relative moisture of approximately72% are consumed by humans and may be processed by canning or freezing.The food uses of maize, in addition to human consumption of maizekernels, include both products of dry- and wet-milling industries. Theprincipal products of maize dry milling are grits, meal and flour. Themaize wet-milling industry can provide maize starch, maize syrups, anddextrose for food use. Maize oil is recovered from maize germ, which isa by-product of both dry- and wet-milling industries.

Maize, including both grain and non-grain portions of the plant, is alsoused extensively as livestock feed, primarily for beef cattle, dairycattle, hogs, and poultry. Industrial uses of maize include productionof ethanol, maize starch in the wet-milling industry and maize flour inthe dry-milling industry. The industrial applications of maize starchand flour are based on functional properties, such as viscosity, filmformation, adhesive properties, and ability to suspend particles. Themaize starch and flour have application in the paper and textileindustries. Other industrial uses include applications in adhesives,building materials, foundry binders, laundry starches, explosives,oil-well muds, and other mining applications. Plant parts other than thegrain of maize are also used in industry: for example, stalks and husksare made into paper and wallboard and cobs are used for fuel and to makecharcoal.

The seed of the present invention or of the present invention furthercomprising one or more single gene traits, the plant produced from theinbred seed, the hybrid maize plant produced from the crossing of theinbred, hybrid seed, and various parts of the hybrid maize plant can beutilized for human food, livestock feed, and as a raw material inindustry.

The present invention therefore also discloses an agricultural productcomprising a plant of the present invention or derived from a plant ofthe present invention. The present invention also discloses anindustrial product comprising a plant of the present invention orderived from a plant of the present invention. The present inventionfurther discloses methods of producing an agricultural or industrialproduct comprising planting seeds of the present invention, growingplant from such seeds, harvesting the plants and processing them toobtain an agricultural or industrial product.

A deposit of at least 2500 seeds of this invention will be maintained bySyngenta Seed Inc. Access to this deposit will be available during thependency of this application to the Commissioner of Patents andTrademarks and persons determined by the Commissioner to be entitledthereto upon request. All restrictions on availability to the public ofsuch material will be removed upon issuance of a granted patent of thisapplication by depositing at least 2500 seeds of this invention at theAmerican Type Culture Collection (ATCC), at 10801 University Boulevard,Manassas, Va. 20110. The ATCC number of the deposit is PTA-12698. Thedate of deposit was Mar. 23, 2012 and the seed was tested on Apr. 9,2012 and found to be viable. The deposit of at least 2500 seeds will befrom inbred seed taken from the deposit maintained by Syngenta Seed Inc.The ATCC deposit will be maintained in that depository, which is apublic depository, for a period of 30 years, or 5 years after the lastrequest, or for the enforceable life of the patent, whichever is longer,and will be replaced if it becomes nonviable during that period.

Additional public information on patent variety protection may beavailable from the PVP Office, a division of the U.S. Government.Accordingly, the present invention has been described with some degreeof particularity directed to the embodiment of the present invention. Itshould be appreciated, though that the present invention is defined bythe following claims construed in light of the prior art so thatmodifications or changes may be made to the embodiment of the presentinvention without departing from the inventive concepts containedherein.

1. A seed of the maize inbred line NPFA4734, representative seed of saidline having been deposited under ATCC Accession Number PTA-12698.
 2. Amaize plant or plant part produced by growing the seed of claim
 1. 3. AnF1 hybrid maize seed produced by crossing a plant of maize inbred lineNPFA4734 of claim 2 with a different maize plant and harvesting theresultant F1 hybrid maize seed.
 4. A maize plant or plant part producedby growing the F1 hybrid maize seed of claim
 3. 5. An F1 hybrid maizeseed comprising an inbred maize plant cell of inbred maize lineNPFA4734, representative seed of said line having been deposited underATCC Accession Number PTA-12698.
 6. A maize plant produced by growingthe F1 hybrid maize seed of claim
 5. 7. Pollen of a maize plant producedby growing the maize seed of claim
 1. 8. A process of introducing adesired trait into maize inbred line NPFA4734 comprising: (a) crossingNPFA4734 plants grown from NPFA4734 seed, representative seed of whichhas been deposited under ATCC Accession Number PTA-12698, with plants ofanother maize line that comprise a desired trait to produce F1 progenyplants, wherein the desired trait is selected from the group consistingof waxy starch, male sterility, herbicide resistance, insect resistance,bacterial disease resistance, fungal disease resistance, and viraldisease resistance; (b) selecting F1 progeny plants that have thedesired trait to produce selected F1 progeny plants; (c) crossing theselected progeny plants with the NPFA4734 plants to produce backcrossprogeny plants; (d) selecting for backcross progeny plants that have thedesired trait to produce selected backcross progeny plants; and (e)repeating steps (c) and (d) at least three or more times to producebackcross progeny plants that comprise the desired trait and all of thephysiological and morphological characteristics of corn inbred lineNPFA4734 listed in Table 1 when grown in the same environmentalconditions.
 9. A plant produced by the process of claim
 8. 10. A maizeplant having all the physiological and morphological characteristics ofinbred line NPFA4734, wherein a sample of the seed of inbred lineNPFA4734 was deposited under ATCC Accession Number PTA-12698.
 11. Aprocess of producing maize seed, comprising crossing a first parentmaize plant with a second parent maize plant, wherein one or both of thefirst or the second parent maize plants is the plant of claim 10, andharvesting the resultant seed.
 12. The maize seed produced by theprocess of claim
 11. 13. The maize seed of claim 12, wherein the maizeseed is hybrid seed.
 14. A hybrid maize plant, or its parts, produced bygrowing said hybrid seed of claim
 13. 15. The maize plant of claim 10,further comprising a genome comprising at least one transgene or a geneconversion conferred by a transgene.
 16. The maize plant of claim 15,wherein the gene confers a trait selected from the group consisting ofherbicide tolerance; insect tolerance; resistance to bacterial, fungal,nematode and viral disease; waxy starch; male sterility or restorationof male fertility; modified carbohydrate metabolism and modified fattyacid metabolism.
 17. A method of producing a maize plant derived fromthe inbred line NPFA4734, the method comprising the steps of (a) growinga progeny plant produced-from a cross comprising the plant of claim 10with a second maize plant; (b) crossing the progeny plant with itself ora different plant to produce a seed of a progeny plant of a subsequentgeneration; (c) growing a progeny plant of a subsequent generation fromsaid seed and crossing the progeny plant of a subsequent generation withitself or a different plant; and (d) repeating steps (b) and (c) for anadditional 0-5 generations to produce a maize plant derived from theinbred line NPFA4734.
 18. A method for developing a maize plant in amaize plant breeding program, comprising applying plant breedingtechniques to the maize plant of claim 10, or its parts, whereinapplication of said techniques results in development of a maize plant.19. The method for developing a maize plant in a maize plant breedingprogram of claim 18, wherein the plant breeding techniques are selectedfrom the group consisting of recurrent selection, backcrossing, pedigreebreeding, restriction fragment length polymorphism enhanced selection,genetic marker enhanced selection, and transformation.